Human negative elongation factor activates transcription and regulates alternative transcription initiation

Jianlong Sun, Rong Li

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

The human negative elongation factor (NELF) is a four-subunit protein complex that inhibits the movement of RNA polymerase II (RNAPII) at an early elongation stage in vitro. NELF-mediated stalling of RNAPII also attenuates transcription of a number of inducible genes in human cells. To obtain a genome-wide understanding of human NELF-mediated transcriptional regulation in vivo, we carried out an exon array study in T47D breast cancer cells with transient small interfering RNA knockdown of individual NELF subunits. Upon depletion of NELF-A, -C, or -E, the vast majority of NELF-regulated genes were down-regulated. Many of the down-regulated genes encode proteins that play key roles in cell cycle progression. Consequently, NELF knockdown resulted in significant reduction in DNA synthesis and cell proliferation. Chromatin immunoprecipitation showed that NELF knockdown led to dissociation of RNAPII from the promoter-proximal region of the cell cycle-regulating genes. This was accompanied by decreased histone modifications associated with active transcription initiation (H3K9Ac) and elongation (H3K36Me3), as well as reduced recruitment of the general transcription factor TFIIB and increased overall histone occupancy at a subset of the down-regulated promoters. Lastly, our study indicates that NELF regulates alternative transcription initiation of BSG (Basigin) gene by differentially influencing RNAPII density at the two neighboring exons at the 5′ end of the gene. Taken together, our data suggest a diverse transcriptional consequence of NELF-mediated RNAPII pausing in the human genome.

Original languageEnglish (US)
Pages (from-to)6443-6452
Number of pages10
JournalJournal of Biological Chemistry
Volume285
Issue number9
DOIs
StatePublished - Feb 26 2010

Fingerprint

Transcription
Genes
RNA Polymerase II
Cells
Histones
Elongation
Exons
CD147 Antigens
Transcription Factor TFIIB
negative elongation factor
General Transcription Factors
Histone Code
cdc Genes
Protein Subunits
Cell proliferation
Chromatin Immunoprecipitation
Human Genome
Small Interfering RNA
Chromatin
Genetic Promoter Regions

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Human negative elongation factor activates transcription and regulates alternative transcription initiation. / Sun, Jianlong; Li, Rong.

In: Journal of Biological Chemistry, Vol. 285, No. 9, 26.02.2010, p. 6443-6452.

Research output: Contribution to journalArticle

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